Avaliação do potencial contra câncer colorretal e da embriotoxicidade em peixe-zebra de um glicosídeo enônico

Abstract

Colorectal cancer (CRC), which affects the large intestine (colon and rectum), is the most common cancer of the gastrointestinal tract and ranks third in incidence, also being responsible for the second highest number of cancer deaths worldwide. In this context, the search for new drugs to combat the disease is a strategic priority for global health, given the public health problem that CRC represents. From this perspective, substances such as glycosides have been studied, and some have shown promising anticancer activity against certain types of the disease. The present study aimed to evaluate the cytotoxicity of a synthetic steroidal glycoside (RO-OX04) in colorectal cancer cell lines and non-tumor cells, predict the pharmacokinetic characteristics of the compound through an insilico study (ADMET), and evaluate its embryotoxicity in an assay with zebrafish (Danio rerio). Initially, in the cytotoxicity assays, obtained through the MTT assay, the following cell lines were used: CRC (HT-29, LoVo and HCT-116) and non-tumor (HEK-293). The compound RO-OX04 proved to be more effective against the LoVo cell line, with IC50 values of 8 μM (24h) and 3.4 μM (72h). Importantly, RO- OX04 showed lower toxicity to non-tumor cells (HEK-293), with IC50 values of 15 μM (24h) and 12 μM (72h). Regarding the type of cell death, the compound induced apoptosis in cells exposed to RO-OX04 (8 μM) within 24h, with a percentage of 30.45 ± 4.5% of the cells in the sample. Concerning its effect on cell migration, the compound reduced groove closure when exposed to 8 μM, with closure percentages of 16 ± 3.5% and 22.2 ± 6% at 24 and 48 h, respectively. Regarding the inhibition of colony formation, exposure to 4, 8, and 12 μM of RO- OX04 significantly inhibited colony formation (35 ± 9, 1 ± 1, and 0 colonies), indicating a strong anticancer effect. The ADMET study predicted a promising pharmacological profile for RO- OX04, with good solubility and oral bioavailability. Furthermore, no neurotoxicity, mutagenicity, carcinogenicity, inhibition of cytochrome p450 isoforms, or interaction with P-glycoprotein were predicted. Furthermore, embryotoxicity analysis revealed that the compound RO-OX04 causes lethality and embryotoxicity in zebrafish after 96 hours of exposure, in a concentration- dependent manner. The results of this work suggest that the compound RO-OX04 is a promising cytotoxic agent for the treatment of colorectal cancer, with a favorable pharmacological profile, justifying the continuation of studies for its development as a future drug.